Simultaneous targeting of NPC1 and VDAC1 by itraconazole leads to synergistic inhibition of MTOR signaling and angiogenesis

Sarah A. Head, Wei Q. Shi, Eun Ju Yang, Benjamin A. Nacev, Sam Y. Hong, Kalyan K. Pasunooti, Ruo Jing Li, Joong Sup Shim, Jun O. Liu

Research output: Contribution to journalArticle

Abstract

The antifungal drug itraconazole was recently found to exhibit potent antiangiogenic activity and has since been repurposed as an investigational anticancer agent. Itraconazole has been shown to exert its antiangiogenic activity through inhibition of the mTOR signaling pathway, but the molecular mechanism of action was unknown. We recently identified the mitochondrial protein VDAC1 as a target of itraconazole and a mediator of its activation of AMPK, an upstream regulator of mTOR. However, VDAC1 could not account for the previously reported inhibition of cholesterol trafficking by itraconazole, which was also demonstrated to lead to mTOR inhibition. In this study, we demonstrate that cholesterol trafficking inhibition by itraconazole is due to direct inhibition of the lysosomal protein NPC1. We further map the binding site of itraconazole to the sterol-sensing domain of NPC1 using mutagenesis, competition with U18666A, and molecular docking. Finally, we demonstrate that simultaneous AMPK activation and cholesterol trafficking inhibition leads to synergistic inhibition of mTOR, endothelial cell proliferation, and angiogenesis.

Original languageEnglish (US)
Pages (from-to)174-182
Number of pages9
JournalACS chemical biology
Volume12
Issue number1
DOIs
StatePublished - 2017

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

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    Head, S. A., Shi, W. Q., Yang, E. J., Nacev, B. A., Hong, S. Y., Pasunooti, K. K., Li, R. J., Shim, J. S., & Liu, J. O. (2017). Simultaneous targeting of NPC1 and VDAC1 by itraconazole leads to synergistic inhibition of MTOR signaling and angiogenesis. ACS chemical biology, 12(1), 174-182. https://doi.org/10.1021/acschembio.6b00849